Study on active capacity and detergent application potential of low-temperature alkaline serine protease produced by new strain Exiguobacterium indicum 1.2.3

Abstract

Microorganisms have long captivated researchers for their potential to produce enzymes with diverse industrial applications. Efficient production of proteases from new strains is crucial as these enzymes play a vital role in breaking down protein bonds, enabling their use in industrial applications. Therefore, a novel Exiguobacterium indicum 1.2.3 was isolated (Istanbul, Turkiye) and characterized in this study. This strain produced alkaline serine protease, which works in lower temperatures (20–40 °C) with casein as a specific substrate. The protease was utterly stable for 3 h at 30 °C. The enzyme was also highly stable in the pH range of 8–11. The optimum activity was obtained at pH 10. The crude enzyme activity was enhanced by various metal ions and retained 147%, 125%, 124%, and 117% of its activity within 1 mM Ca²⁺, Mn²⁺, Cu²⁺, and Mg²⁺, respectively. The crude enzyme was inactive with phenylmethylsulfonyl fluoride, indicating a serine residue on the active side. The enzyme exhibited a significant proteolytic effect in the presence of surfactants and oxidizing agents. The addition of Tween 80, Triton X-100, and sodium perborate improved enzymatic activity up to 135%, 109%, and 105%, respectively. According to the washing results, the crude enzyme effectively removed the blood on different types of standard pre-stained textiles at 30 °C. In conclusion, Exiguobacterium indicum 1.2.3 is a promising candidate for protease production, with its diverse applications spanning various industrial sectors, particularly detergents.

Keywords

Exiguobacterium indicum / Alkaline protease / Low-temperature detergent / Industrial enzymes / Enzymatic activity

References

Anbu P, Annadurai G, Hur BK. Production of alkaline protease from a newly isolated Exiguobacterium profundum BK-P23 evaluated using the response surface methodology. Biologia, 2013

Beg QK, Gupta R. Purification and characterization of an oxidation-stable, thiol-dependent serine alkaline protease from Bacillus mojavensis. Enzyme Microbial Technol, 2003

Ben Bacha A, Al-Assaf A, Moubayed NMS, Abid I. Evaluation of a novel thermo-alkaline Staphylococcusaureus lipase for application in detergent formulations. Saudi J Biol Sci, 2018, 25(3): 409-417.

Chen X, Wang L, Zhou J, Wu H, Li D, Cui Y, Lu B. Exiguobacterium sp A1b/GX59 isolated from a patient with community-acquired pneumonia and bacteremia: genomic characterization and literature review. BMC Infect Dis, 2017

Collins MD, Lund BM, Farrow JAE, Schleifer KH. Chemotaxonomic study of an Alkalophilic Bacterium Exiguobacteriumaurantiacum gen nov sp nov. Microbiology, 1983

Degering C, Eggert T, Puls M, Bongaerts J, Evers S, Maurer KH, Jaeger KE. Optimization of protease secretion in bacillus subtilis and bacillus licheniformis by screening of homologousand heterologous signal peptides. Appl Environ Microbiol, 2010

Emon TH, Hakim A, Chakraborthy D, Bhuyan FR, Iqbal A, Hasan M, Aunkor TH, Azad AK. Kinetics, detergent compatibility and feather-degrading capability of alkaline protease from Bacillussubtilis AKAL7 and Exiguobacteriumindicum AKAL11 produced with fermentation of organic municipal solid wastes. J Environ Sci Health Part A Toxic/hazardous Substances Environ Eng, 2020

Ghorbel S, Kammoun M, Soltana H, Nasri M, Hmidet N. Streptomycesflavogriseus HS1: Isolation and characterization of extracellular proteases and their compatibility with laundry detergents. BioMed Res Int, 2014

Grbavčić S, Bezbradica D, Izrael-Živković L, Avramović NŠ, Milosavić N, KaradŽić I, Knezevic-Jugovic Z. Production of lipase and protease from an indigenous Pseudomonasaeruginosa strain and their evaluation as detergent additives: compatibility study with detergent ingredients and washing performance. Biores Technol, 2011, 102(24): 11226-11233.

Hakim A, Bhuiyan FR, Iqbal A, Emon TH, Ahmed J, Azad AK. Production and partial characterization of dehairing alkaline protease from Bacillus subtilis AKAL7 and Exiguobacterium indicum AKAL11 by using organic municipal solid wastes. Heliyon, 2018

Hammami A, Hamdi M, Abdelhedi O, Jridi M, Nasri M, Bayoudh A. Surfactant- and oxidant-stable alkaline proteases from Bacillus invictae: characterization and potential applications in chitin extraction and as a detergent additive. Int J Biol Macromol, 2017, 96: 272-281.

Hammami A, Fakhfakh N, Abdelhedi O, Nasri M, Bayoudh A. Proteolytic and amylolytic enzymes from a newly isolated Bacillusmojavensis SA: characterization and applications as laundry detergent additive and in leather processing. Int J Biol Macromol, 2018, 108: 56-68.

Kumar, S. S., Jithin, V., Jijeesh, V., Gayathri, V., Shiburaj, S., Haridas, M., and Sabu, A. (2018). Production and purification of alkaline protease from Exiguobacterium indicum TBG-PICH-001 isolated from soil samples of Pichavaram Estuary (Tamil Nadu). Indian Journal of Geo-Marine Sciences, 47(3).

Laitala K, Jensen HM. Cleaning effect of household laundry detergents at low temperatures. Tenside Surf Detergents, 2010

Li Q, Yi L, Marek P, Iverson BL. Commercial proteases: present and future. FEBS Lett, 2013

Maruthiah T, Esakkiraj P, Prabakaran G, Palavesam A, Immanuel G. Purification and characterization of moderately halophilic alkaline serine protease from marine Bacillus subtilis AP-MSU 6. Biocatal Agric Biotechnol, 2013, 2(2): 116-119.

Oberoi R, Beg QK, Puri S, Saxena RK, Gupta R. Characterization and wash performance analysis of an SDS-stable alkaline protease from a Bacillus sp. World J Microbiol Biotechnol, 2001, 17(5): 493-497.

Olajuyigbe FM, Falade AM. Purification and partial characterization of serine alkaline metalloprotease from bacillus brevis MWB-01. Bioresourc Bioprocess, 2014

Otto BR, Van Dooren SJM, Nuijens JH, Luirink J, Oudega B. Characterization of a hemoglobin protease secreted by the pathogenic Escherichiacoli strain EB1. J Experim Med, 1998

Raveendran S, Parameswaran B, Ummalyma SB, Abraham A, Mathew AK, Madhavan A, Rebello S, Pandey A. Applications of microbial enzymes in food industry. Food Technol Biotechnol, 2018

Sani JT, Omolbanin S, Gharibi S, Shariati MA. The importance of alkaline protease commercial applications: a short review. Indian J Res Pharm Biotechnol, 2017, 5(1): 5.

Senthilkumar SR, Ashokkumar B, Chandra Raj K, Gunasekaran P. Optimization of medium composition for alkali-stable xylanase production by Aspergillusfischeri Fxn 1 in solid-state fermentation using central composite rotary design. Biores Technol, 2005, 96(12): 1380-1386.

Sun F, Hu Y, Chen Q, Kong B, Liu Q. Purification and biochemical characteristics of the extracellular protease from Pediococcuspentosaceus isolated from Harbin dry sausages. Meat Sci, 2019

Suwannaphan S, Fufeungsombut E, Promboon A, Chim-anage P. A serine protease from newly isolated Bacillus sp. for efficient silk degumming, sericin degrading and colour bleaching activities. Int Biodeterior Biodegradation, 2017, 117: 141-149.

Vojcic L, Pitzler C, Körfer G, Jakob F, Martinez R, Maurer KH, Schwaneberg U. Advances in protease engineering for laundry detergents. In New Biotechnol, 2015

Zhang D, Zhu Z, Li Y, Li X, Guan Z, Zheng J. Comparative genomics of exiguobacterium reveals what makes a cosmopolitan bacterium. Msystems, 2021

Zhang Y, Hu J, Zhang Q, Cai D, Chen S, Wang Y. Enhancement of alkaline protease production in recombinant Bacilluslicheniformis by response surface methodology. Bioresourc Bioprocess, 2023